法医学杂志

法医学杂志 ›› 2016, Vol. 32 ›› Issue (5): 342-345.DOI: 10.3969/j.issn.1004-5619.2016.05.006

• 论著 • 上一篇    下一篇

多重置换扩增在含抑制物检材中的应用

丁东雪，丁  梅   

  1. （中国医科大学法医学院法医血清教研室，辽宁 沈阳 110001）
  • 发布日期:2016-10-25 出版日期:2016-10-28
  • 通讯作者: 丁梅，女，教授，博士研究生导师，主要从事法医物证学研究；E-mail：dingmei1209@gmail.com
  • 作者简介:丁东雪（1988—），女，硕士研究生，主要从事法医物证学研究；E-mail：dingdongxue1@hotmail.com

Application of Multiple Displacement Amplification in Samples with Inhibitors

DING DONG-XUE, DING MEI   

  1. （Department of Forensic Serology, Faculty of Forensic Medicine, China Medical University, Shenyang 110001, China）
  • Online:2016-10-25 Published:2016-10-28

摘要： 目的 研究多重置换扩增（multiple displacement amplification，MDA）在含抑制物检材中的抗抑制能力，与磁珠法纯化检材相比较，证明其在法医学中的应用及意义。 方法 将不同浓度血红素和腐殖酸与样本DNA进行混合，分为MDA处理组、磁珠法处理组和空白对照组，PCR-STR单基因座D3S1358扩增联合聚丙烯酰胺凝胶电泳检测，并应用AmpF?詛STR?誖IdentifilerTM Plus试剂盒联合毛细管电泳检测。 结果 血红素质量浓度大于1 ng/μL或腐殖酸质量浓度大于0.1 ng/μL时，空白对照组经单基因座STR检测不能得到扩增产物；磁珠法处理组血红素质量浓度大于100 ng/μL或腐殖酸浓度大于1 ng/μL时，不能够得到扩增产物；MDA处理组各浓度抑制物均能成功扩增，完全不受抑制物影响。 结论 MDA技术可消除血红素及腐殖酸的抑制作用，其抗抑制能力优于磁珠法纯化DNA，具有一定的法医学应用意义。

关键词: 法医遗传学, 核酸扩增技术, 多重置换扩增, 抑制物, 血红素, 腐殖酸

Abstract: Objective To explore the ability of inhibition resistibility of multiple displacement amplification （MDA） in samples with inhibitors. To explain the application and value of MDA in forensic medicine by comparing with using magnetic beads methods （MBM） to purify sample. Methods Different concentrations of hemoglobin and humid acid （HA） mixed with DNA samples and then divided the samples into MDA group, MBM group and control group. D3S1358 locus was amplified and detected by polyacrylamide gel electrophoresis detection system and AmpF?詛STR?誖IdentifilerTM Plus Kit-capillary electrophoresis detection system. Results When hemoglobin concentrations exceed 1 ng/μL or HA concentrations exceed 0.1 ng/μL, amplification products could not be obtained by single-locus system in control group. When hemoglobin concentration exceeds 100 ng/μL or HA concentrations exceed 1 ng/μL, the samples could not be amplified by MBM. Inhibitors in different concentrations were amplified successfully in MDA group without any influence from inhibitors. Conclusion MDA has the capability to remove the inhibition of hemoglobin and HA, which is better than MBM and has a certain value in forensic practices.

Key words: forensic genetics, nucleic acid amplification techniques, multiple displacement amplification, inhibitors, hemoglobin, humic acid

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